This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Koreen, L. Articles by Kreiswirth, B. N. Search for Related Content PubMed PubMed Citation Articles by Koreen, L. Articles by Kreiswirth, B. N.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, August 2005, p. 3985-3994, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.3985-3994.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Comparative Sequencing of the Serine-Aspartate Repeat-Encoding Region of the Clumping Factor B Gene (clfB) for Resolution within Clonal Groups of Staphylococcus aureus

Larry Koreen,^1,2 Srinivas V. Ramaswamy,^3, Steven Naidich,⁴ Irina V. Koreen,¹ Gavin R. Graff,⁵ Edward A. Graviss,³ and Barry N. Kreiswirth^2*

New Jersey Medical School and Graduate School of Biomedical Sciences, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103,¹ Public Health Research Institute, International Center for Public Health, Newark, New Jersey 07103,² Department of Pathology, Baylor College of Medicine, Houston, Texas 77030,³ eGenomics, New York, New York 10013,⁴ Department of Pediatrics, Penn State College of Medicine, Hershey, Pennsylvania 17033⁵

Received 23 December 2004/ Returned for modification 12 February 2005/ Accepted 16 March 2005

Molecular techniques such as spa typing and multilocus sequence typing use DNA sequence data for differentiating Staphylococcus aureus isolates. Although spa typing is capable of detecting both genetic micro- and macrovariation, it has less discriminatory power than the more labor-intensive pulsed-field gel electrophoresis (PFGE) and costly genomic DNA microarray analyses. This limitation hinders strain interrogation for newly emerging clones and outbreak investigations in hospital or community settings where robust clones are endemic. To overcome this constraint, we developed a typing system using DNA sequence analysis of the serine-aspartate (SD) repeat-encoding region within the gene encoding the keratin- and fibrinogen-binding clumping factor B (clfB typing) and tested whether it is capable of discriminating within clonal groups. We analyzed 116 S. aureus strains, and the repeat region was present in all isolates, varying in sequence and in length from 420 to 804 bp. In a sample of 36 well-characterized genetically diverse isolates, clfB typing subdivided identical spa and PFGE clusters which had been discriminated by whole-genome DNA microarray mapping. The combination of spa typing and clfB typing resulted in a discriminatory power (99.5%) substantially higher than that of spa typing alone and closely approached that of the whole-genome microarray (100.0%). clfB typing also successfully resolved genetic differences among isolates differentiated by PFGE that had been collected over short periods of time from single hospitals and that belonged to the most prevalent S. aureus clone in the United States. clfB typing demonstrated in vivo, in vitro, and interpatient transmission stability yet revealed that this locus may be recombinogenic in a primarily clonal population structure. Taken together, these data show that the SD repeat-encoding region of clfB is a highly stable marker of microvariation, that in conjunction with spa typing it may serve as a DNA sequence-based alternative to PFGE for investigating genetically similar strains, and that it is useful for analyzing collections of isolates in both long-term population-based and local epidemiologic studies.

Present address: Channing Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.

This article has been cited by other articles:

• Balajee, S. A., Tay, S. T., Lasker, B. A., Hurst, S. F., Rooney, A. P. (2007). Characterization of a Novel Gene for Strain Typing Reveals Substructuring of Aspergillus fumigatus across North America. Eukaryot Cell 6: 1392-1399 [Abstract] [Full Text]  
• Tormo, M. A., Ubeda, C., Marti, M., Maiques, E., Cucarella, C., Valle, J., Foster, T. J., Lasa, I., Penades, J. R. (2007). Phase-variable expression of the biofilm-associated protein (Bap) in Staphylococcus aureus. Microbiology 153: 1702-1710 [Abstract] [Full Text]  
• Monk, A. B., Archer, G. L. (2007). Use of Outer Surface Protein Repeat Regions for Improved Genotyping of Staphylococcus epidermidis. J. Clin. Microbiol. 45: 730-735 [Abstract] [Full Text]  
• Kuhn, G., Francioli, P., Blanc, D. S. (2007). Double-Locus Sequence Typing Using clfB and spa, a Fast and Simple Method for Epidemiological Typing of Methicillin-Resistant Staphylococcus aureus. J. Clin. Microbiol. 45: 54-62 [Abstract] [Full Text]  
• Strommenger, B., Kettlitz, C., Weniger, T., Harmsen, D., Friedrich, A. W., Witte, W. (2006). Assignment of Staphylococcus Isolates to Groups by spa Typing, SmaI Macrorestriction Analysis, and Multilocus Sequence Typing.. J. Clin. Microbiol. 44: 2533-2540 [Abstract] [Full Text]